Measurement of Transient Permeability of Sp2/0 Myeloma Cells: Flow Cytometric Study

Novickij, Vitalij; Girkontaitė, Irutė; Grainys, Audrius; Zinkevičienė, Auksė; Lastauskienė, Eglė; Švedienė, Jurgita; Paškevičius, Algimantas; Markovskaja, Svetlana; Novickij, Jurij

doi:10.1515/msr-2016-0038

Cited by 13 publications

(5 citation statements)

References 28 publications

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“…Later, the cells (after pulsing) were transferred to 1.5 mL tubes (Eppendorf, Hamburg, Germany) for analysis by the FlowSight (Amnis, Seattle, WA, USA) flow cytometer. Gate definition and processing were performed in accordance with a previous study [ 53 ].…”

Section: Methodsmentioning

confidence: 99%

Electrochemotherapy Using Doxorubicin and Nanosecond Electric Field Pulses: A Pilot in Vivo Study

et al. 2020

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Pulsed electric field (PEF) is frequently used for intertumoral drug delivery resulting in a well-known anticancer treatment—electrochemotherapy. However, electrochemotherapy is associated with microsecond range of electrical pulses, while nanosecond range electrochemotherapy is almost non-existent. In this work, we analyzed the feasibility of nanosecond range pulse bursts for successful doxorubicin-based electrochemotherapy in vivo. The conventional microsecond (1.4 kV/cm × 100 µs × 8) procedure was compared to the nanosecond (3.5 kV/cm × 800 ns × 250) non-thermal PEF-based treatment. As a model, Sp2/0 tumors were developed. Additionally, basic current and voltage measurements were performed to detect the characteristic conductivity-dependent patterns and to serve as an indicator of successful tumor permeabilization both in the nano and microsecond pulse range. It was shown that nano-electrochemotherapy can be the logical evolution of the currently established European Standard Operating Procedures for Electrochemotherapy (ESOPE) protocols, offering better energy control and equivalent treatment efficacy.

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Section: Methodsmentioning

confidence: 99%

Electrochemotherapy Using Doxorubicin and Nanosecond Electric Field Pulses: A Pilot in Vivo Study

et al. 2020

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“…The samples of the cell suspension (63 μL) were mixed with 7 μL propidium iodide (PI) (Sigma-Aldrich, Germany) at final concentration of 50 μM [51] and transferred to electroporation cuvette for pulse application. After the pulsing, a 10-min incubation at room temperature was performed for passive dye diffusion, followed by flow cytometric analysis.…”

Section: Methodsmentioning

confidence: 99%

“…The samples were analyzed by FlowSight (Amnis, Seattle, USA) flow cytometer. The fluorescent cells (PI permeable) were gated as permeabilized in accordance with standard gating strategy used in electroporation studies [51–53].…”

Section: Methodsmentioning

confidence: 99%

Low concentrations of acetic and formic acids enhance the inactivation of Staphylococcus aureus and Pseudomonas aeruginosa with pulsed electric fields

et al. 2019

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Background Skin infections, particularly caused by drug-resistant pathogens, represent a clinical challenge due to being a frequent cause of morbidity and mortality. The objectives of this study were to examine if low concentrations of acetic and formic acids can increase sensitivity of Staphylococcus aureus and Pseudomonas aeruginosa to pulsed electric field (PEF) and thus, promote a fast and efficient treatment methodology for wound treatment. Results We have shown that the combination of PEF (10–30 kV/cm) with organic acids (0.1% formic and acetic acids) increased the bactericidal properties of treatment. The effect was apparent for both acids. The proposed methodology allowed to reduce the energy of electrical pulses and the inhibitory concentrations of acids, while still maintain high efficiency of bacteria eradication. Conclusions Application of weak organic acids as bactericidal agents has many advantages over antibiotics because they do not trigger development of drug-resistance in bacteria. The combination with PEF can make the treatment effective even against biofilms. The results of this study are particularly useful for the development of new methodologies for the treatment of extreme cases of wound infections when the chemical treatment is no longer effective or hinders wound healing.

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“…PI fluorescence was evaluated using bandpass filter of 610–630 nm. The fluorescent cells (PI permeable) were gated as permeabilized in accordance with typical gating strategies used in electroporation studies (Michie et al., 2000; Novickij et al., 2016a, 2018a). A shift of fluorescence spectra and the cells in the defined gate (which was defined based on the untreated control) have been interpreted as fluorescence positive (permeabilized), while the cells outside the gate have been interpreted as nonfluorescent (nonpermeabilized).…”

Section: Methodsmentioning

confidence: 99%

Inactivation of Escherichia coli Using Nanosecond Electric Fields and Nisin Nanoparticles: A Kinetics Study

et al. 2018

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Nisin is a recognized bacteriocin widely used in food processing, however, being ineffective against gram-negative bacteria and in complex food systems. As a result, the research of methods that have cell wall–permeabilizing activity is required. In this study, electroporation to trigger sensitization of gram-negative bacteria to nisin-loaded pectin nanoparticles was used. As a model microorganism, bioluminescent strain of E. coli was introduced. Inactivation kinetics using nanosecond pulsed electric fields (PEFs) and nisin nanoparticles have been studied in a broad range (100–900 ns, 10–30 kV/cm) of pulse parameters. As a reference, the microsecond range protocols (100 μs × 8) have been applied. It was determined that the 20–30 kV/cm electric field with pulse duration ranging from 500 to 900 ns was sufficient to cause significant permeabilization of E. coli to trigger a synergistic response with the nisin treatment. The kinetics of the inactivation was studied with a time resolution of 2.5 min, which provided experimental evidence that the efficacy of nisin-based treatment can be effectively controlled in time using PEF. The results and the proposed methodology for rapid detection of bacteria inactivation rate based on bioluminescence may be useful in the development and optimization of protocols for PEF-based treatments.

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Measurement of Transient Permeability of Sp2/0 Myeloma Cells: Flow Cytometric Study

Cited by 13 publications

References 28 publications

Electrochemotherapy Using Doxorubicin and Nanosecond Electric Field Pulses: A Pilot in Vivo Study

Electrochemotherapy Using Doxorubicin and Nanosecond Electric Field Pulses: A Pilot in Vivo Study

Low concentrations of acetic and formic acids enhance the inactivation of Staphylococcus aureus and Pseudomonas aeruginosa with pulsed electric fields

Inactivation of Escherichia coli Using Nanosecond Electric Fields and Nisin Nanoparticles: A Kinetics Study

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